Activating Compound | Comment | Organism | Structure |
---|---|---|---|
50S ribosome | stimulates the enzyme | Escherichia coli | |
70S ribosome | required, stimulates the enzyme | Escherichia coli | |
additional information | the 30S ribosome does not stimulate the enzyme | Escherichia coli |
Cloned (Comment) | Organism |
---|---|
gene lepA, recombinant overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
H81A | site-directed mutagenesis | Escherichia coli |
additional information | generation of C-terminal domain truncation mutants of the enzyme, truncation of the C-terminal domain does not abolish ribosome binding, the mutant enzyme is stabilized on the ribosome when the C-terminal domain is removed. Binding of wild-type EF4 and mutant variants to the ribosome in the presence of guanine nucleotides, kinetics and affinities | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Thiostrepton | inhibits the stimulation of the enzyme's GTPase activity by the 70S ribosome | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | Michaelis-Menten kinetics of GTP hydrolysis, stopped flow measurements, kinetic analysis of wild-type and mutant enzymes, overview | Escherichia coli | |
0.00032 | - |
GTP | recombinant wild-type enzyme in presence of 70S ribosome, pH 7.5, 37°C | Escherichia coli | |
0.27 | - |
GTP | recombinant wild-type enzyme in absence of 70S ribosome, pH 7.5, 37°C | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + H2O | Escherichia coli | - |
GDP + phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P60785 | gene lepA | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chhrmatography and gel filtration | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
GTP + H2O | - |
Escherichia coli | GDP + phosphate | - |
? | |
additional information | enzyme-ribosome binding analysis, overview. Binding of wild-type EF4 and mutant variants to the ribosome in the presence of guanine nucleotides, kinetics and affinities | Escherichia coli | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
EF4 | - |
Escherichia coli |
elongation factor 4 | - |
Escherichia coli |
LepA | - |
Escherichia coli |
ribosome-dependent GTPase | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
GTP | recombinant wild-type enzyme in absence of 70S ribosome, pH 7.5, 37°C | Escherichia coli | |
2.3 | - |
GTP | recombinant wild-type enzyme in presence of 70S ribosome, pH 7.5, 37°C | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
General Information | Comment | Organism |
---|---|---|
malfunction | successive removal of the C-terminus impairs ribosome-dependent multiple turnover GTPase activity of enzyme EF4 | Escherichia coli |
additional information | the last 44 C-terminal amino acids of elongation factor 4 form a subdomain within the C-terminal domain that is important for GTP-dependent function on the ribosome. Efficient nucleotide hydrolysis by the enzyme on the ribosome depends on its conserved residue His 81, which is essential for catalysis in EF4 | Escherichia coli |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
GTP | recombinant wild-type enzyme in presence of 70S ribosome, pH 7.5, 37°C | Escherichia coli | |
0.011 | - |
GTP | recombinant wild-type enzyme in absence of 70S ribosome, pH 7.5, 37°C | Escherichia coli |